Amplite® Fluorimetric Sphingomyelinase Assay Kit *Red Fluorescence*

Amplite® Fluorimetric Sphingomyelinase Assay Kit

Red Fluorescence

Five types of sphingomyelinase (SMase) have been identified based on their cation dependence and pH optima of action. They are lysosomal acid SMase, secreted zinc-dependent acid SMase, magnesium-dependent neutral SMase, magnesium-independent neutral SMase, and alkaline SMase. Among the five types, the lysosomal acidic SMase and the magnesium-dependent neutral SMase are considered major candidates for the production of ceramide in the cellular response to stress. Our Amplite® Fluorimetric Sphingomyelinase Assay Kit provides the most sensitive method for detecting neutral SMase activity or screening its inhibitors. The kit uses Amplite® Red as a fluorogenic probe to indirectly quantify the phosphocholine produced from the hydrolysis of sphingomyelin (SM) by sphingomyelinase (SMase). It can be used for measuring the SMase activity in blood, cell extracts or other solutions. The fluorescence intensity of Amplite® Red is proportional to the formation of phosphocholine, therefore to the SMase activity. Amplite® Red enables the assay readable either in fluorescence intensity or absorption mode. The kit is an optimized "mix and read" assay that can be used for real time monitoring of Smase activities. Our kit 13622 has been developed for monitoring acid SMase activity.

Sphingomyelinase dose response was measured on a solid black 96-well plate with Amplite® Fluorimetric Sphingomyelinase Assay Kit using a Gemini fluorescence microplate reader (Molecular Devices).

Protocol

SDS

COA

Catalog	Size	Price	Quantity
13621	200 Tests	Price

Citations

View all 6 citations: Citation Explorer

Doxepin mitigates noise induced neuronal damage in primary auditory cortex of mice via suppression of acid sphingomyelinase/ceramide pathway

Authors:

Su, Yu-Ting and Meng, Xing-Xing and Zhang, Xi and Guo, Yi-Bin and Zhang, Hai-Jun and Cheng, Yao-Ping and Xie, Xiao-Ping and Chang, Yao-Ming and Bao, Jun-Xiang

Journal:

The Anatomical Record (2017)

Riccardin DN induces lysosomal membrane permeabilization by inhibiting acid sphingomyelinase and interfering with sphingomyelin metabolism in vivo

Authors:

Li, Lin and Niu, Huanmin and Sun, Bin and Xiao, Yanan and Li, Wei and Yuan, Huiqing and Lou, Hongxiang

Journal:

Toxicology and Applied Pharmacology (2016): 175--184

New Aspects of Silibinin Stereoisomers and their 3-O-galloyl Derivatives on Cytotoxicity and Ceramide Metabolism in Hep G2 hepatocarcinoma Cell Line

Authors:

Mashhadi Akbar Boojar, Mahdi and Ejtemaei Mehr, Shahram and Hassanipour, Mahsa and Mashhadi Akbar Boojar, Masoud and Dehpour, Ahmad Reza

Journal:

Iranian Journal of Pharmaceutical Research (2016): 421--433

Mitochondrial respiration controls lysosomal function during inflammatory T cell responses

Authors:

Baixauli, Francesc and Acín-Pérez, Rebeca and Villarroya-Beltrí, Carolina and Mazzeo, Carla and Nunez-Andrade, Norman and Gab, undefined and é-Rodriguez, Enrique and Ledesma, Maria Dolores and Blázquez, Alberto and Martin, Miguel Angel and Falcón-Pérez, Juan Manuel and others, undefined

Journal:

Cell metabolism (2015): 485--498

The ATP-binding cassette transporter-2 (ABCA2) regulates esterification of plasma membrane cholesterol by modulation of sphingolipid metabolism

Authors:

Davis, Warren

Journal:

Biochimica et Biophysica Acta (BBA)-Molecular and Cell Biology of Lipids (2014): 168--179

References

View all 65 references: Citation Explorer

A novel mitochondrial sphingomyelinase in zebrafish cells

Authors:

Yabu T, Shimuzu A, Yamashita M.

Journal:

J Biol Chem (2009): 20349

A novel sphingomyelinase-like enzyme in Ixodes scapularis tick saliva drives host CD4 T cells to express IL-4

Authors:

Alarcon-Chaidez FJ, Boppana VD, Hagymasi AT, Adler AJ, Wikel SK.

Journal:

Parasite Immunol (2009): 210

Alterations of myelin-specific proteins and sphingolipids characterize the brains of acid sphingomyelinase-deficient mice, an animal model of Niemann-Pick disease type A

Authors:

Buccinna B, Piccinini M, Prinetti A, Sc and roglio F, Prioni S, Valsecchi M, Votta B, Grifoni S, Lupino E, Ramondetti C, Schuchman EH, Giordana MT, Sonnino S, Rinaudo MT.

Journal:

J Neurochem (2009): 105

A complex extracellular sphingomyelinase of Pseudomonas aeruginosa inhibits angiogenesis by selective cytotoxicity to endothelial cells

Authors:

Vasil ML, Stonehouse MJ, Vasil AI, Wadsworth SJ, Goldfine H, Bolcome RE, 3rd, Chan J.

Journal:

PLoS Pathog (2009): e1000420

Expression of alkaline sphingomyelinase in yeast cells and anti-inflammatory effects of the expressed enzyme in a rat colitis model

Authors:

Andersson D, Kotarsky K, Wu J, Agace W, Duan RD.

Journal:

Dig Dis Sci (2009): 1440

Spectral properties

Excitation (nm)	571
Emission (nm)	584

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
UNSPSC	12352200

Instrument settings

Fluorescence microplate reader
Excitation	540 nm
Emission	590 nm
Cutoff	570 nm
Recommended plate	Solid black

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Page updated on October 13, 2025

Component A: Enzyme Mix	2 bottles (lyophilized powder)
Component B: Sphingomyelin	1 vial (100 µL)
Component C: Amplite™ Red	1 vial (lyophilized powder)
Component D: SMase Reaction Buffer	1 bottle (10 mL)
Component E: Assay Buffer	1 bottle (20 mL)
Component F: Sphingomyelinase Standard	1 vial (0.2 unit, lyophilized powder)
Component G: DMSO	1 vial (200 µL)